Limits...
Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometric (LC/ESI-MS/MS) Study for the Identification and Characterization of In Vivo Metabolites of Cisplatin in Rat Kidney Cancer Tissues: Online Hydrogen/Deuterium (H/D) Exchange Study.

Bandu R, Ahn HS, Lee JW, Kim YW, Choi SH, Kim HJ, Kim KP - PLoS ONE (2015)

Bottom Line: A total of thirty one unknown in vivo metabolites have been identified and the structures of metabolites were elucidated using LC-MS/MS experiments combined with accurate mass measurements.Online HDX experiments have been used to further support the structural characterization of metabolites.The results showed that CP undergoes a series of ligand exchange biotransformation reactions with water and other nucleophiles like thio groups of methionine, cysteine, acetylcysteine, glutathione and thioether.

View Article: PubMed Central - PubMed

Affiliation: Department of Applied Chemistry, College of Applied Sciences, Kyung Hee University, Yong-in City, Republic of Korea.

ABSTRACT
In vivo rat kidney tissue metabolites of an anticancer drug, cisplatin (cis-diamminedichloroplatinum [II]) (CP) which is used for the treatment of testicular, ovarian, bladder, cervical, esophageal, small cell lung, head and neck cancers, have been identified and characterized by using liquid chromatography positive ion electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) in combination with on line hydrogen/deuterium exchange (HDX) experiments. To identify in vivo metabolites, kidney tissues were collected after intravenous administration of CP to adult male Sprague-Dawley rats (n = 3 per group). The tissue samples were homogenized and extracted using newly optimized metabolite extraction procedure which involves liquid extraction with phosphate buffer containing ethyl acetate and protein precipitation with mixed solvents of methanol-water-chloroform followed by solid-phase clean-up procedure on Oasis HLB 3cc cartridges and then subjected to LC/ESI-HRMS analysis. A total of thirty one unknown in vivo metabolites have been identified and the structures of metabolites were elucidated using LC-MS/MS experiments combined with accurate mass measurements. Online HDX experiments have been used to further support the structural characterization of metabolites. The results showed that CP undergoes a series of ligand exchange biotransformation reactions with water and other nucleophiles like thio groups of methionine, cysteine, acetylcysteine, glutathione and thioether. This is the first research approach focused on the structure elucidation of biotransformation products of CP in rats, and the identification of metabolites provides essential information for further pharmacological and clinical studies of CP, and may also be useful to develop various effective new anticancer agents.

No MeSH data available.


Related in: MedlinePlus

Proposed fragmentation mechanism for metabolites M11-M14.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4526507&req=5

pone.0134027.g009: Proposed fragmentation mechanism for metabolites M11-M14.

Mentions: M11 ([M+H]+; m/z 572.0922): The metabolite M11 at m/z 572.0922 ([M+H]+; C10H24N5O6PtSCl; -2.11 ppm) was eluted at 8.2 min. The appearance of diagnostic product ion at m/z 264.4732 due to loss of C10H18N3O6S (Fig 9) in the LC-MS/MS spectrum of protonated M11, which is also a characteristic product ion observed in the LC-MS/MS of protonated CP, M1, M4, M5, M7 and M9, authenticates the presence of Pt+(NH3)2(Cl) moiety in its structure. The protonated M11 yields diagnostic product ion at m/z 308.0916 corresponding to protonated glutathione, confirms the presence of glutathione moiety in its structure. This was further confirmed by MS/MS of deuterated M11 in which the m/z 308.0916 ion got shifted to m/z 316.1426 due to incorporation of eight deuterium atoms (-NH2, two-NH, two-COOH,-SH and mobile proton). The LC-MS/MS spectrum of protonated M11 also shows other structure indicative fragment ions at m/z 526.1466 (loss of HCOOH), m/z 443.0811 (loss of C5H7NO3), m/z 323.1365 (simultaneous elimination of C3H5NO3 & C5H10N2O3) and m/z 130.0491 (C5H8NO3+) which are consistent with the proposed structure for M11. Based on these data, M11 was identified as a glutathione metabolite of CP.


Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometric (LC/ESI-MS/MS) Study for the Identification and Characterization of In Vivo Metabolites of Cisplatin in Rat Kidney Cancer Tissues: Online Hydrogen/Deuterium (H/D) Exchange Study.

Bandu R, Ahn HS, Lee JW, Kim YW, Choi SH, Kim HJ, Kim KP - PLoS ONE (2015)

Proposed fragmentation mechanism for metabolites M11-M14.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4526507&req=5

pone.0134027.g009: Proposed fragmentation mechanism for metabolites M11-M14.
Mentions: M11 ([M+H]+; m/z 572.0922): The metabolite M11 at m/z 572.0922 ([M+H]+; C10H24N5O6PtSCl; -2.11 ppm) was eluted at 8.2 min. The appearance of diagnostic product ion at m/z 264.4732 due to loss of C10H18N3O6S (Fig 9) in the LC-MS/MS spectrum of protonated M11, which is also a characteristic product ion observed in the LC-MS/MS of protonated CP, M1, M4, M5, M7 and M9, authenticates the presence of Pt+(NH3)2(Cl) moiety in its structure. The protonated M11 yields diagnostic product ion at m/z 308.0916 corresponding to protonated glutathione, confirms the presence of glutathione moiety in its structure. This was further confirmed by MS/MS of deuterated M11 in which the m/z 308.0916 ion got shifted to m/z 316.1426 due to incorporation of eight deuterium atoms (-NH2, two-NH, two-COOH,-SH and mobile proton). The LC-MS/MS spectrum of protonated M11 also shows other structure indicative fragment ions at m/z 526.1466 (loss of HCOOH), m/z 443.0811 (loss of C5H7NO3), m/z 323.1365 (simultaneous elimination of C3H5NO3 & C5H10N2O3) and m/z 130.0491 (C5H8NO3+) which are consistent with the proposed structure for M11. Based on these data, M11 was identified as a glutathione metabolite of CP.

Bottom Line: A total of thirty one unknown in vivo metabolites have been identified and the structures of metabolites were elucidated using LC-MS/MS experiments combined with accurate mass measurements.Online HDX experiments have been used to further support the structural characterization of metabolites.The results showed that CP undergoes a series of ligand exchange biotransformation reactions with water and other nucleophiles like thio groups of methionine, cysteine, acetylcysteine, glutathione and thioether.

View Article: PubMed Central - PubMed

Affiliation: Department of Applied Chemistry, College of Applied Sciences, Kyung Hee University, Yong-in City, Republic of Korea.

ABSTRACT
In vivo rat kidney tissue metabolites of an anticancer drug, cisplatin (cis-diamminedichloroplatinum [II]) (CP) which is used for the treatment of testicular, ovarian, bladder, cervical, esophageal, small cell lung, head and neck cancers, have been identified and characterized by using liquid chromatography positive ion electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) in combination with on line hydrogen/deuterium exchange (HDX) experiments. To identify in vivo metabolites, kidney tissues were collected after intravenous administration of CP to adult male Sprague-Dawley rats (n = 3 per group). The tissue samples were homogenized and extracted using newly optimized metabolite extraction procedure which involves liquid extraction with phosphate buffer containing ethyl acetate and protein precipitation with mixed solvents of methanol-water-chloroform followed by solid-phase clean-up procedure on Oasis HLB 3cc cartridges and then subjected to LC/ESI-HRMS analysis. A total of thirty one unknown in vivo metabolites have been identified and the structures of metabolites were elucidated using LC-MS/MS experiments combined with accurate mass measurements. Online HDX experiments have been used to further support the structural characterization of metabolites. The results showed that CP undergoes a series of ligand exchange biotransformation reactions with water and other nucleophiles like thio groups of methionine, cysteine, acetylcysteine, glutathione and thioether. This is the first research approach focused on the structure elucidation of biotransformation products of CP in rats, and the identification of metabolites provides essential information for further pharmacological and clinical studies of CP, and may also be useful to develop various effective new anticancer agents.

No MeSH data available.


Related in: MedlinePlus